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WO1999045769A2 - Solution - Google Patents

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Publication number
WO1999045769A2
WO1999045769A2 PCT/GB1999/000711 GB9900711W WO9945769A2 WO 1999045769 A2 WO1999045769 A2 WO 1999045769A2 GB 9900711 W GB9900711 W GB 9900711W WO 9945769 A2 WO9945769 A2 WO 9945769A2
Authority
WO
WIPO (PCT)
Prior art keywords
cell
preservation
preservation solution
tissue
organ
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/GB1999/000711
Other languages
English (en)
Other versions
WO1999045769A3 (fr
Inventor
Christoph Berwanger
Gerry Stansby
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Imperial College of London
Original Assignee
Imperial College of London
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from GBGB9805049.5A external-priority patent/GB9805049D0/en
Priority claimed from GBGB9813572.6A external-priority patent/GB9813572D0/en
Application filed by Imperial College of London filed Critical Imperial College of London
Priority to AU27382/99A priority Critical patent/AU2738299A/en
Publication of WO1999045769A2 publication Critical patent/WO1999045769A2/fr
Publication of WO1999045769A3 publication Critical patent/WO1999045769A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/10—Preservation of living parts
    • A01N1/12—Chemical aspects of preservation
    • A01N1/122—Preservation or perfusion media
    • A01N1/125—Freeze protecting agents, e.g. cryoprotectants or osmolarity regulators
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/10—Preservation of living parts

Definitions

  • the present invention relates to a solution.
  • the present invention relates to a solution that is useful in preserving, such as maintaining for long periods of time, organs, tissues and cells.
  • UW solution is widely used in transplantation and its composition is:
  • the present invention seeks to provide a preservation solution that is able to preserve tissues and organs for long periods of time.
  • a preservation solution for preserving a cell wherein the preservation solution comprises at least D 2 O; and wherein the cell is a vascular cell.
  • a method of preserving a cell comprising contacting the cell with a preservation solution such that preservation of the cell occurs; wherein the preservation solution comprises at least D 2 O; and wherein the cell is a vascular cell.
  • a preservation solution to preserve a cell; wherein the preservation solution comprises at least D 2 O; and wherein the cell is a vascular cell.
  • preservation solutions for cells which lead to better preservation times etc.
  • preservation occurs for at least 12 hours, more preferably for at least 24 hours, more preferably for at least 36 hours, more preferably for at least 48 hours, more preferably for at least 60 hours, more preferably for at least 72 hours.
  • H 2 O preservation solutions that are to be used for preserving cells, such as isolated cells or cells contained within organ or tissue, and in doing so derive beneficial effects.
  • D 2 O deuterium oxide
  • H 2 O water
  • D 2 O has been investigated for use in preserving liver, kidney and pancreas for transplantation (4, 5, 6).
  • experimental work has yielded differing results with D 2 O showing benefits over water in kidney preservation, whilst having no effects on pancreas preservation (4, 6) and deleterious effects on heart preservation.
  • D 2 O acts on cells by stabilising biological membranes - since deuterium bonds are stronger that hydrogen bonds - and/or by lowering intracellular calcium levels - which are usually raised during hypothermia.
  • the breakdown of calcium regulation within the cell that is associated with cooling is well documented and believed to play a vital role in the damage that occurs to a cell at low temperatures (3).
  • high cytosolic calcium promotes the activation of degradative phospholipase (e.g. phospholipase A 2 ) and protease enzymes which break down cellular proteins and membranes (11).
  • a further aspect of the present invention is the use of D 2 O in the manufacture of a preservation solution to stabilise a biological membrane of a cell and/or to lower intracellular calcium levels of a cell.
  • the present invention also provides a method of stabilising a biological membrane of a cell and/or lowering intracellular calcium levels of a cell, the method comprising contacting the cell with a preservation solution comprising D 2 O such that stabilisation of a biological membrane of a cell occurs and/or lowering intracellular calcium levels of a cell occurs.
  • the present invention also provides a method of improving the viability of the endothelial layer during the preservation of a tissue or an organ, wherein the method comprises contacting the tissue or organ with a preservation solution comprising D 2 O such that of improvement of the viability of the endothelial layer occurs.
  • the present invention also provides the use of D 2 O to reduce platelet aggregation and/or thrombus formation and/or to ensure production of vasodilators layer during the preservation of a tissue or an organ. Otherwise expressed the present invention also provides a method of reducing platelet aggregation and/or thrombus formation and/or to ensure production of vasodilators during the preservation of a tissue or an organ, wherein the method comprises contacting the tissue or organ with a preservation solution comprising D 2 O such that reduction of platelet aggregation and/or thrombus formation and/or ensurement of production of vasodilators occurs.
  • the present invention also provides the use of D 2 O to inhibit deterioration of vascular tissue in an organ that is to be used for transplantation.
  • cell as used herein with reference to the present invention means a cell when alone or when present in a tissue or organ.
  • the term means a cell when present in a tissue or organ.
  • the cell is a vascular cell.
  • the tissue is preferably a blood vessel.
  • the blood vessel is a human internal mammary artery or a human saphenous vein.
  • the preservation solution is preferably suitable for preserving the cell under cryogenic conditions.
  • the term "cell” means a cell when present in an organ
  • the organ is preferably a kidney, more preferably a human kidney.
  • the preservation solution is preferably suitable for preserving the cell under hypothermic conditions.
  • a preservation solution for preserving a cell wherein the preservation solution comprises at least D 2 O; wherein the cell is a vascular cell; and wherein the cell is present in a human kidney.
  • the preservation solution of the present invention may contain conventional ingredients - such as any one or more of salts, biocidal agents, anti-oxidants, rheology modifiers (such as thinners), stabilisers, colourings (such as dyes), sugars, starches, proteins, enzymes, chelants, amino acids, nutrients, etc.
  • the solvent of the preservation solution of the present invention may contain other solvents - such as any one or more of H 2 O or other suitable solvent which may be organic or inorganic in nature.
  • the preservation solution of the present invention may be prepared by mixing at some stage one or more ingredients in D 2 O.
  • the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 9: 1 to 1 :9.
  • the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 8:1 to 1:8.
  • the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 7:1 to 1:7. More preferably, the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 6:1 to 1:6.
  • the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 5:1 to 1:5.
  • the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 4:1 to 1 :4.
  • the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 3: 1 to 1:3.
  • the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 2:1 to 1:3.
  • the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 1:1 to 1:3.
  • the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of about 1 :3.
  • the solvent of the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 9:1 to 1:9.
  • the solvent of the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 8:1 to 1:8.
  • the solvent of the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 7 : 1 to 1 :7. More preferably, the solvent of the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 6:1 to 1:6.
  • the solvent of the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 5:1 to 1:5.
  • the solvent of the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 4:1 to 1:4.
  • the solvent of the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 3:1 to 1:3.
  • the solvent of the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 2:1 to 1:3.
  • the solvent of the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O and H 2 O of from 1:1 to 1:3.
  • the solvent of the preservation solution comprises D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of about 1:3.
  • the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of from 9:1 to 1:9.
  • the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of from 8:1 to 1:8.
  • the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of from 7:1 to 1:7.
  • the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of from 6: 1 to 1:6.
  • the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of from 5:1 to 1:5.
  • the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of from 4: 1 to 1:4.
  • the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of from 3:1 to 1:3.
  • the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of from 2:1 to 1:3.
  • the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of from 1:1 to 1:3.
  • the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of about 1:3. Otherwise expressed, the solvent of the preservation solution consists of about 25% D 2 O and about 75% H 2 O.
  • the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of 1:3. Otherwise expressed, the solvent of the preservation solution consists of 25% D 2 O and 75% H 2 O.
  • the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D O to H 2 O of from 3:1 to 1:0. Otherwise expressed, the solvent of the preservation solution consists of from 75 % D 2 O and 25 %
  • the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of from 9:1 to 1:0. Otherwise expressed, the solvent of the preservation solution consists of from 90% D 2 O and 10% H 2 O to 100% D 2 O and 0% H 2 O. In a yet more preferred embodiment, the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of from 19:1 to 1:0. Otherwise expressed, the solvent of the preservation solution consists of from 95% D 2 O and 5% H 2 O to 100% D 2 O and 0% H 2 O.
  • the solvent of the preservation solution consists of D 2 O and H 2 O, which are in relative amounts of D 2 O to H 2 O of from 1:0. Otherwise expressed, the solvent of the preservation solution consists of from 100% D 2 O and 0% H 2 O. These preferred ratios are particularly preferred wherein the preservation solution is to preserve a cell under cryogenic conditions.
  • the preservation solution of the present invention wherein the solvent of the preservation solution consists of from 75% D 2 O and 25% H 2 O to 100% D 2 O and 0% H 2 O, allows cells to be preserved for longer periods of time under cryogenic conditions.
  • This increased preservation is compared to preservation with 100% H 2 O or with prior art preservation solutions.
  • the increase in preservation is particularly marked, wherein the solvent of the preservation solution of the present invention consists of 100% D 2 O and 0% H 2 O.
  • the preservation solution of the present invention wherein the solvent of the preservation solution consists of from 25 % D 2 O and 75% H 2 O to 100% D 2 O and 0% H 2 O, allows cells to be preserved for longer periods of time under hypothermic conditions.
  • the preservation solution of the present invention allows cells to be preserved for longer periods of time. Otherwise expressed, we believe that proportionate replacement with D 2 O in accordance with the present invention may extend preservation times. In particular, we have found that preferred solutions of the present invention can significantly (p ⁇ 0.05) improve the viability (such as endothelial and/or smooth muscle cell function) of vascular tissue up to about 72 hours or even more.
  • Figure 1 shows the ability of tissue containing a vascular cell to contract after 24 hours storage in a preservation solution.
  • Figure 2 shows the ability of tissue containing a vascular cell to contract after 48 hours storage in a preservation solution.
  • Figure 3 shows the ability of tissue containing a vascular cell to contract after 72 hours storage in a preservation solution.
  • Figure 4 shows the ability of tissue containing a vascular cell to relax after 24 hours storage in a preservation solution.
  • Figure 5 shows the ability of tissue containing a vascular cell to relax after 48 hours storage in a preservation solution.
  • Figure 6 shows the ability of tissue containing a vascular cell to relax after 72 hours storage in a preservation solution.
  • composition of KH is:
  • H100 - which is normal UW solution - i.e. lactobionic acid lOOmM, KOH 100 ⁇ 5mM, allopurinal ImM, KH 2 PO 4 25mM, MgSO 4 Heptahydrate 5mM, raffinose 30mM, glutathione 3mM, NaOH 25 ⁇ 5mM, adenosine 5mM (but omitting hydroxyethyl starch) in 100% H 2 O.
  • D25 - which is the same as H100 except that the H 2 O solvent is now 25% D 2 O/75%
  • D100 - which is the same as H100 except that the H 2 O solvent is now 100% D 2 O (D100).
  • D 2 O was obtained from Aldrich (Gillingham), Dorset, UK), all other chemicals from Sigma (Poole, Dorset, UK).
  • the aortic segments were stored in these preservation solutions at 4°C for 24, 48 or 72h.
  • rat aortic segments were stored in UW-solutions based on 100% H 2 O, 25% D 2 O, 50% D 2 O and 100% D 2 O at 4°C for 24, 48 or 72h.
  • Vascular function was measure via contraction and endothellum-dependent relaxation following stimulation with phenylephrine and acetylcholine.
  • the present invention is not limited to D 2 O containing UW solutions.
  • any suitable preservation solution may be used - providing the preservation solution comprises D 2 O.
  • the present invention shows that it is possible to replace (partially or completely) H 2 O with D 2 O in solutions that are to be used as organ, tissue or cell preservation solutions. We believe that the present invention will have important implications in a number of medical fields.
  • the present invention will have an important use in the transplantation of all types of solid organs - such as one or more of heart, lung, liver, pancreas and kidney.
  • the vasculature is the interface between the graft and the host.
  • the present invention will have use in preserving cells or tissues or organs of a vascular nature.
  • the present invention will have use in preserving different cells or tissues or organs - i.e. other than those of a vascular nature.
  • cryopreservation - which involves freezing and storing cells (such as tissues and organs) at temperatures as low as -196°C or even lower.
  • D 2 O in a preservation solution could ameliorate the adverse effects of freezing cells.
  • D 2 O in accordance with the present invention could also be used to improve allografts (such as implantation of vascular tissue) or xenografts (such as implantation of porcine pancreatic islets). This would result in an improvement in the treatment of, for example, peripheral vascular disease or diabetes.
  • D 2 O in accordance with the present invention could also be used to improve the storage of cell lines for research or commercial purposes.
  • D 2 O in accordance with the present invention could also be used to improve the storage of other cell types - such as oocyctes and/ or sperms and/ or embryos.
  • adenosine (mM/L) 5 5 allopurinol (mM/L) 1 1 glucose (mM/L) 139 glutathione (mM/L) 3 3 histidine (mM/L) 180 histidine HC1.H 2 0 (mM/L) 18 a-ketoglutaric acid (mM/L) 1 lactobionic acid (mM/L) 100 100 mannitol (mM/L) 30 raffinose (mM/L) 30 30 tryptophan (mM/L) 2
  • FISCHER JH, JESCHKEIT S Effectivity of freshly prepared or refreshed solutions for heart preservation versus commercial Eurocollins, Bretschneider's HTK, or University of Wisconsin solution. Transplantation 1995; 59:1259- 1262.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Dentistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Environmental Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention porte sur une solution préservatrice de cellules du type vasculaire comprenant au moins du D2O.
PCT/GB1999/000711 1998-03-10 1999-03-10 Solution Ceased WO1999045769A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU27382/99A AU2738299A (en) 1998-03-10 1999-03-10 Solution

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
GB9805049.5 1998-03-10
GBGB9805049.5A GB9805049D0 (en) 1998-03-10 1998-03-10 Solution
GB9813572.6 1998-06-24
GBGB9813572.6A GB9813572D0 (en) 1998-06-24 1998-06-24 Solution

Publications (2)

Publication Number Publication Date
WO1999045769A2 true WO1999045769A2 (fr) 1999-09-16
WO1999045769A3 WO1999045769A3 (fr) 1999-11-04

Family

ID=26313261

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1999/000711 Ceased WO1999045769A2 (fr) 1998-03-10 1999-03-10 Solution

Country Status (2)

Country Link
AU (1) AU2738299A (fr)
WO (1) WO1999045769A2 (fr)

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2253086C3 (de) * 1972-10-25 1979-02-15 Martin Prof. Dr. 1000 Berlin Wenzel Konservierende Lösungen zur Infusion und zur Aufbewahrung von Geweben und Gewebeteilen
DE2516836A1 (de) * 1975-04-15 1976-10-28 Boehringer Sohn Ingelheim Loesungen oder medien fuer die tieftemperatur-lagerung von biologischem material

Also Published As

Publication number Publication date
AU2738299A (en) 1999-09-27
WO1999045769A3 (fr) 1999-11-04

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